Executive summary SBIR award (1R43NS107079-01): The neuronal ceroid-lipofuscinoses (NCLs) are lysosomal storage diseases (LSDs) with a prevalence of approximately 1.5 to nine per million population (1.3 to 7 per 100,000 live births). The infantile onset form termed CLN1 disease is characterized by progressive intellectual and motor deterioration, seizures, loss of vision, and early death. This is caused by mutations in the CLN1 gene, which codes for the lysosomal enzyme palmitoyl-protein thioesterase-1 (PPT1) resulting in a reduction or absence of enzyme activity. CLN1 disease usually presents between 6 and 24 months of age and there are 2-3 children with this form identified each year and 24 known children with CLN1 in the US (with likely many more undiagnosed). There is no treatment available. Collaborations Pharmaceuticals, Inc. is working with Dr. Jonathan Cooper and colleagues at the Washington University of St. Louis (WUSTL) to test an enzyme replacement therapy for human recombinant PPT1 by combined intracerebroventricular and intrathecal dosing in PPT1 knockout mice. Aim 1. Production and Characterization of rhPPT1. We will produce sufficient quantities of rhPPT1 and characterize the enzyme activity Aim 2. Intracerebroventricular and intrathecal administration of rhPPT1. With the recent finding of significant spinal cord pathology that precedes brain pathology in Cln1 mice, and that therapeutically targeting this region improves the pathology and extends lifespan and motor performance, we propose treating Cln1 mice with monthly doses of rhPPT1 by either ICV or IT injections alone or by a combination of these routes and test for their effects, if any, on enzyme activity, gait abnormalities and histopathology. If we are successful in demonstrating that a combination of intracerebroventricular and intrathecal delivery of rhPPT1 can improve the neuropathology, motor performance and restore enzyme activity, in Phase II we will pursue efficacy studies in a larger animal model and perform IND enabling toxicology (immunotoxicology). An effective enzyme replacement for PPT1-related NCL would become the standard of care as no treatment is currently available. Further, the development of an ERT would be complimentary to any efforts to develop gene therapy for this disease. Our potential return on investment for this treatment would be obtaining the FDA rare pediatric disease priority review voucher. Progress to date: We have initiated Aim 1 to make additional enzyme (enough for large animal studies) using CHO DG44 cells and have set up our lab at Collaborations Pharmaceuticals Inc. to do cell culture and protein development for rare diseases. We have initiated Aim 2 to perform the in vivo studies at WUSTL. To date there have been no major technical, administrative, or commercial challenges. We have also initiated discussions with clinical research organizations and contract manufacturing organizations (e.g. Calvert labs and MassBiologics) to obtain quotes for future toxicology studies and making protein, respectively. I-Corps Team: (CEO/PI) Dr. Sean Ekins is the CEO of Collaborations Pharmaceuticals, Inc. (CPI) and also the PI of this grant. He brings more than 23 years of preclinical Drug Metabolism and Pharmacokinetics (DMPK), toxicology, and drug discovery research in large pharmaceutical, biotechnology, informatics software and several start-up virtual drug companies and other companies (e.g. Collaborative Drug Discovery). He has obtained funding for >20 NIH/DOD grants as the PI and has worked extensively on drug discovery. As CEO of CPI since founding it in 2016 he has been responsibility for the scientific success of research projects, working closely with collaborators to develop projects, providing project management, scientific marketing and commercialization. CPI have been awarded 9 grants to date from the NIH and DOD. He is therefore actively involved in several grants and collaborative projects working on drug discovery for rare and neglected diseases. His goal is to industrialize drug discovery for rare diseases and demonstrate that treatments can be developed cost effectively by anyone. (PI/Senior Scientist) Dr. Ana Puhl Rubio is the senior scientist working closely on this grant at CPI and collaborating with the team at Washington University of St. Louis. She brings extensive knowledge and technical expertise in protein expression and purification, biochemistry and biophysical techniques relevant to drug discovery and translational research. She has completed 3 Postdocs, made many proteins and solved 22 protein structures. She has also attended numerous business classes and courses for life science PhDs. (Business Advisory Board member/ Industry expert) Andrea Barry (BA, MS) is the President of Eleventh Hour Inc. She has over 20 years of successful leadership and management experience in sales and marketing, with a focus on startups and visionary product releases. (MDL, Scitegic, Accelrys, Vertical*i and CDD). Andrea has been very closely involved with CPI since it was founded as a member of the board and she has provided business guidance, consulting and coaching to the CEO. Each team member is committed to the time requirements of the program.